Histidine containing nutriceutical

ABSTRACT

Nutriceutical compositions useful as a dietary supplement which have antioxidant/free radical scavengers and also having a cytoprotective effect are disclosed. The compositions contain a cytoprotective and antioxidant/free radical scavenging amount of at least one of D-histidine, L-histidine, a racemic mixture thereof, a non-racemic mixture thereof, and nutriceutically acceptable salts thereof in combination with phytonutrients having antioxidant properties such as canthanaxin, vitamin A and limonene. The compositions can be prepared in capsule form, tablets, sustained release tablets, suspensions and oral rehydration solutions.

This application is a continuation of application Ser. No. 08/962,655filed Nov. 3, 1997 which application is now U.S. Pat. No. 5,972,985.

FIELD OF THE INVENTION

The present invention relates to compositions of matter in the field ofnutriceuticals, phytochemicals, and phytonutrients. More in particular,the present investigation relates to dietary supplements whichincorporate cytoprotective agents, free radical scavengers andantioxidants. The instant invention is also directed to antioxidantformulations and vitamin supplements which incorporate histidine as anantioxidant, as a free radical scavenger and as a cytoprotective agent.The invention also provides compositions that scavenge singlet oxygen.The present invention also provides sustained release formulationscontaining histidine as well as histidine in combination withantioxidants and other nutrients and micronutrients. The instantinvention also provides a method for providing nutrition to a mammal inneed thereof by administering histidine in combination with othervitamin supplements.

BACKGROUND OF THE INVENTION

When free radicals attack biological cells and tissues they can beincredibly destructive. They damage our cells, which can mean prematureaging, reduced immune function, inflammation and ultimately degenerativedisease. Among our primary defenses are the antioxidant nutrients, ofwhich the most well known are vitamins C and E. The biological activityof antioxidants is enormously important to our health. Antioxidants havebeen known to neutralize free radical damage, rendering them all butharmless. Aging and deterioration in human cells is caused by thechemical process of oxidation. Scientists theorize that when pollutants,chemicals, and toxins such as cigarette smoke or food additives combinewith oxygen in your bloodstream, they produce unstable (and harmful)molecules called “free radicals”. Free radicals cause cells to die offfaster than your body can produce new ones. As you get older, your bodyproduces fewer new cells to replace the ones that die. This combinationresults in the aging process. When free radicals attack human cells,they weaken cell walls and erode them to the core. These weakened cellsage rapidly, inviting disease and infection because of their unhealthystate. Scientists have identified a group of vitamins calledantioxidants which choke off the supply of oxygen to the free radicals,thereby rendering them almost harmless. With free radicals thusneutralized, human cells and tissue remain younger and healthier.

The term nutriceutical appears to have been introduced initially byStephen de Felice, M.D., director of New York's Foundation forInnovation in Medicine. The term nutriceutical is intended to describespecific chemical compounds found in foods that may prevent disease. Theterm phytochemical is a more recent evolution of the term thatemphasizes the plant source of most of these protective,disease-preventing compounds. A true nutritional role for phytochemicalsis becoming more probable every day as research uncovers more of theirremarkable benefits. In fact, the term phytonutrient better describesthe compounds' status. Someday, phytochemicals may indeed be classifiedas essential nutrients. A significant body of research suggests a stronglink between diet and health. Current studies are showing that as wemove away from the diet of our ancestors we succumb to modern diseases.Evidence of this can be seen in societies such as the centenarian tribesthat live in remote villages in the Andes mountains and who stillembrace traditional dietary practices. These people have been reportedto live extraordinarily long lives that are free of such illnesses ascancer, heart disease and arthritis. Since few people in our modernsociety live today as do the tribes in remote Andean villages,researchers have examined epidemiological evidence from modern societiesfor clues to the diet-disease connection. On the basis of such studies,biochemical researchers have identified certain phytochemicals that aidthe body in maintaining health and combating disease. As an overallguideline, health authorities recommend that we consume diets rich inwhole grains and fresh fruits and vegetables as well as reduce fat andanimal-protein consumption.

The amino acid L-histidine having the chemical structure shown below

is one of the 10 essential amino acids which cannot be synthesized bythe human body and therefore must be obtained by the diet. In additionto histidine's routine metabolic role as a protein building block,histidine is an effective scavenger of toxic oxygen species andtherefore is useful in the protection of cells and tissues from avariety of inflammatory disease processes.

Several in vitro and in vivo laboratory studies have demonstratedhistidine's antioxidant effects. The antioxidant effect observed withhistidine has been attributed to its ability to scavenge highly reactiveoxygen species, particularly singlet oxygen (¹O₂) and hydroxyl radical(.OH ) produced by the degradation of hydrogen peroxide and molecularoxygen. In one study [A. K. Vinnikova et al “Singlet Oxygen-inducedinhibition of cardiac sarcolemmal Na⁺K⁺-ATPase”, J. Mol. Cell. Card.,24: 465-470 (1992) and R. C. Kureja et al “Singlet oxygen interactionwith Ca²⁺⁻ ATPase of cardiac sarcoplasmic reticulum”; Circ. ReS. 69,1003-1014 (1991)], histidine afforded dose-dependent protection tosarcolemmal Na⁺-K⁺-ATPase and Ca²⁺-ATPase subjected to singletoxygen-mediated damage in vitro. The researchers of the above studiescompared the effects of histidine, superoxide dismutase (SOD) andcatalase in sarcoplasmic is reticulum preparations and noted thathistidine was efficient at protecting the enzymes' integrity due to itsunique ability to scavenge singlet oxygen species. In another study [M.A. Khalid et al “Histidine protects bovine endothelial cells againstanoxia reoxygenation injury by scavenging singlet oxygen”; Circulation,86: I223 (1992)] bovine endothelial cells treated with histidine wereprotected from post-anoxic reoxygenation injury.

Other studies [A. K. Vinnikova et al “Singlet Oxygen-induced inhibitionof cardiac sarcolemmal Na⁺K⁺-ATPase”, J. Mol. Cell. Card., 24: 465-470(1992); Q. Cai et al “Antioxidative properties of histidine and itseffect on myocardial injury during ischemia reperfusion in isolated ratheart”, J. Cardiovasc. Pharmacol.; 25: 147-155 (1995); R. C. Kukreja etal “Protective effects of histidine during ischemia-reperfusion inisolated perfused rat hearts”, Am. J. Physiol., 264:H1370-H1381 (1993)and R. C. Kukreja et al “The oxygen free radical system: from equationthrough membrane-protein interactions to cardivascular injury andprotection”, Cardiovasc. Res. 26: 641-655 (1992)] have shown that canineand rodent heart preparations subjected to ischemia/reperfusion injurywere protected from singlet, hydroxyl radical and superoxide aniondamage by the application of L-histidine.

Recent in vivo studies in rats subjected to coronary artery ligation andreperfusion demonstrated that 3 mM histidine could reduce ventricularinfarcts and the duration and severity of arrhythmias, and was moreeffective than superoxide dismutase at achieving this protectant effect.

In applicants pending U.S. application Ser. No. 08/718,705 filed Sep.27, 1996, it is disclosed that histidine has a strong protective effectagainst intestinal fluid accumulation induced by both Salmonellainfection and by cholera toxin in a mouse model. S. typhimurium-inducedintestinal fluid accumulation was reduced 47% in histidine-treated mice,and evaluation of tissues by light and electron microsocopy showedsignificant protection of the intestinal mucosa by histidine. Inaddition, mean cholera toxin-induced intestinal fluid accumulation wasreduced 54% in histidine-treated mice. The fact that these twodiarrhea-inducing agents act by different mechanisms indicates thathistidine's intestinal protective effects may be very broad.

In a study dealing with mechanism of action [M. C. Erikson et al,“Influence of histidine on lipid peroxidation in sarcoplasmicreticulum”; Arch. Biochem. Biophys. 292: 427-432 (1992)], the effects ofhistidine on lipid peroxidation in an in vitro system derived from fishmuscle sarcoplasnic reticulum were shown to be dependent on the order ofaddition of components (histidine, ferric iron, and reducednicotinamide-adenine-dinucleotide). The studies by Erikson et al suggestthat histidine's protective effect in vivo may be related to its abilityto interfere with the reduction of ferric iron. Ginsburg [I. Ginsburg,“Cationic polyelectrolytes: Potent opsonic agents which activate therespiratory burst in leukocytes”; Free Radic. Res. Commun. 8:11-26(1989)] has shown that polyhistidine or histidine complexed to the freeradical scavengers, SOD or catalase, enhanced their ability to functionas intracellular antioxidants. In vitro and in vivo studies [A. A.Boldyrev, “Natural histidine-containing dipeptide carnosine as a potenthydrophilic antioxidant with membrane stabilizing function”; Mol. Chem.Neuropathol., 19: 185-192 (1993) and O. V. Naumova et al; “Effect ofcarnosine on liver enzyme systems of animals subjected to radiation”;Biokhimiya, 57: 1373-1377 (1992] with the dipeptide carnosine(β-Alanine-L-histidine) indicated that its radioprotective and membranestabilizing effects were attributed to the actions of histidine, notalanine. Camosine functions as a pH buffer and as a chelator of avariety of metals, which contributes to its oxygen scavenging profile.

The prior art is silent regarding nutriceuticals and/or dietarysupplements containing histidine as a cytoprotective agent and as anantioxidant/free radical scavenging agent. Furthermore, the prior art issilent on nutriceuticals and/or dietary supplements incorporatinghistidine as a scavenger of singlet oxygen. The present inventionprovides nutriceutical formulations incorporating histidine as anantioxidant/free radical scavenger and therefore fills a long-felt neednot currently available in the market place.

ADVANTAGES OF THE INVENTION

A primary advantage of the present invention is to provide nutraceuticalcompositions incorporating cytoprotective agents.

An additional advantage of the present invention is to providenutraceutical compositions incorporating histidine as a cytoprotectiveagent.

Another important advantage of the present invention is to providehistidine containing antioxidant/free radical scavenging dietarysupplement formulations.

A further advantage of the invention is to provide dietary supplementsincorporating antioxidants in combination with cytoprotective agents.

Another advantage of the present invention is to provide compositionscontaining histidine and antioxidants and being capable of efficientlyscavenging singlet oxygen providing a complementary protective effect.

A still further advantage of the invention is to provide vitaminformulations incorporating histidine as a cytoprotective agent.

An additional advantage of the present invention is to provide sustainedrelease formulations of histidine in combination with antioxidants andvitamins.

A further advantage of the invention is to provide soft gelatin shellvitamin formulations incorporating histidine as a cytoprotective agent.

Other advantages will become apparent as further described below in thepreferred embodiments of the present invention.

SUMMARY OF THE INVENTION

Briefly, the present invention is directed to a nutriceuticalcomposition comprising: (a) an effective amount of a cyto-protectiveagent; and (b) an effective amount of one or more of a group ofphytonutrients having antioxidant activity.

In another aspect, the present invention relates to an antioxidantdietary supplement comprising: (a) an effective amount of histidine; (b)an effective amount of ascorbic acid; and (c) an effective amount ofalpha-tocopherol.

The instant invention is also directed to a dietary supplementcomprising: (a) an effective amount of a synthetic or naturallyoccurring vitamin or mixtures thereof; and (b) an effectivecytoprotective amount of histidine.

The invention also provides a method of providing nutrition to a mammalin need thereof, which method comprises administering to said mammal acomposition comprising: (a) an effective amount of a cyto-protectiveagent wherein said cyto-protective agent is selected from the groupconsisting of at least one of D-histidine, L-histidine, a racemicmixture thereof, a non-racemic mixture thereof, and nutriceuticallyacceptable salts thereof; and (b) an effective amount of at least one ofa phytonutrient having antioxidant activity wherein said phytonutrienthaving antioxidant activity is selected from the group consisting ofcarotenoids, flavonoids, tocopherols, ascorbates and mixtures thereof.

The present invention is also directed to sustained release formulationscomprising cytoprotective agents and antioxidants having speciallydesigned coatings.

The instant invention is further directed to a controlled releaseformulation in tablet form for once-daily oral administration of about500-1500 mg of at least one of D-histidine, L-histidine, a racemicmixture thereof, a non-racemic mixture thereof, and nutriceuticallyacceptable salts thereof, which tablet comprises a homogeneous matrixcomprising: about 3-12 weight percent of hydroxypropyl mehylcellulosehaving a number average molecular weight in the range of from about50,000 to about 150,000; about 75-97 weight percent of at least one ofD-histidine, L-histidine, a racemic mixture thereof, a non-racemicmixture thereof, and nutriceutically acceptable salts thereof in anamount effective for once daily oral administration; and about 0.05 toabout 4 weight percent of a pharmaceutically acceptable lubricatingagent.

DESCRIPTION OF THE PREFERRED EMBODIMENTS

The compositions of the present invention typically contain acytoprotective agent and one or more phytonutrients having antioxidantactivity. The ideal cytoprotective agent as well as antioxidant/freeradical scavenger for use with the compositions of the present inventionis the naturally occurring form of histidine e.g., L-histidine. However,other forms of histidine that can be used in the practice of the presentinvention include D-histidine, a racemic mixture thereof, a non-racemicmixture thereof, as well as pharmaceutically acceptable salts andprodrugs thereof including polyhistidine or short peptides containingeffective releasable amounts of histidine. Histidine is a particularlypreferred component of the nutriceutical of the present inventionbecause it has been shown to protect cells and tissues from damagemediated by the hydroxyl radical (.OH) and singlet oxygen (¹O₂).Histidine is one of only a few agents that effectively scavenge singletoxygen, a highly reactive form of molecular oxygen with an extraunpaired electron which has been shown to be an important cause of ischemical reperfusion injury. Also the favorable pharmacokineticproperties [N. G. Sitton et al, “Kinetic investigations into thepossible cause of low serum histidine in rheumatoid arthritis”; Ann.Rheum. Dis., 47: 48-52 (1988) and D. A. Gerber, “Determination ofhistidine in serum with O-Phthaldialdehyde”, Anal. Biochem., 34:500-504(1970)] of histidine makes its use as an antioxidant free radicalscavenger dietary supplement very attractive since it is rapidly andextensively absorbed, with maximum plasma concentrations reached onehour or less after an oral dose and bioavailability in the range of 80%to 95%.

Other amino acids such as lysine, arginine, and glycine can also be usedin combination with histidine. Of course it is understood that allnaturally occurring forms of the amino acids can be used in addition toother forms such as the D-forms and racemic and non-racemic mixtures.

Another important component of the nutriceutical of the instantinvention are the phytonutrients. The preferred phytonutrients are thosehaving antioxidant activity. For example, terpenes such as those foundin green foods, soy products and grains, comprise one of the largestclasses of phytonutrients. Terpenes of particular interest includecarotenoids such as beta-carotene. The terpenes function asantioxidants, protecting lipids, blood and other body fluids fromassault by free radical oxygen species including singlet oxygen,hydroxyl, peroxide and superoxide radicals. Terpenoids are dispersedwidely throughout the plant kingdom, protecting plants from the samereactive oxygen species that attack human cells.

This terpene subclass consists of bright yellow, orange and red plantpigments found in vegetables such as tomatoes, parsley, oranges, pinkgrapefruit, spinach and red palm oil. Carotenoids also provide thebright colors seen in some animals, for example flamingos owe theircolor to carotenoids, as do shellfish. Egg yolks are yellow because ofcarotenoids that protect the unsaturated fats in the yolk.

The carotenoid family actually includes two distinct types of molecules.One type, the carotenes, are chemically classified as 40-carbontetraterpenes, which do not include specific chemical features likehydroxyl or keto groups. This type of carotenoid includes the familiarmolecule beta carotene. The second type of carotenoids, thexanthophylls, includes the chemical compounds known as the carotenoidalcohols and keto-carotenoids. In this second category are included themolecules zeaxanthin, cryptoxanthin, and astazanthin.

There are more than 600 naturally occurring carotenoids. Among thecarotenes, only alpha, beta and epsilon carotene possess vitamin Aactivity. Of these, beta carotene is the most active. Alpha carotenepossesses 50 percent to 54 percent of the antioxidant activity of betacarotene, whereas epsilon carotene has 42 percent to 50 percent of theantioxidant activity. The above-mentioned carotenes, along with gammacarotene and the carotenes, lycopene, and lutein, which do not convertto vitamin A, seem to offer protection against lung, colorectal, breast,uterine and prostate cancers (A. Bendich et al, FASEB J 3: 1927-32;January 1989).

Carotenes are tissue-specific in their protection. Overall protectiveeffects are therefore greater when all carotenes are taken together.Carotenes are also known to enhance immune response and protect skincells against UV radiation (A. Bendich, J Nutr, 119: 112-5 January1989). Additionally, they spare the glutathionine Phase IIdetoxification enzymes in the liver that we rely on to safely eliminatepollutants and toxins from the body.

The xanthophyll type of carotenoids which can also be used in thepractice of the instant invention include molecules such ascanthaxantin, cryptoxanthin, zeaxanthin and astaxanthin. Xanthophyllsare important because they appear to protect vitamin A, vitamin E andother carotenoids from oxidation. Evidence is also emerging thatxanthophylls are tissue specific. Cryptoxanthin, for example, may behighly protective of vaginal, uterine and cervical tissues (R. S.Parker., J Nutr, 119: 101-4; January 1989).

The limonoids comprise another terpene subclass which can be used in theinventive compositions. The limonoids are typically found in citrusfruit peels, and their usefulness appears to be linked to protection oflung tissue. In one study, a standardized extract of d-limonene, pinene,and eucalyptol was effective in clearing congestive mucus from the lungsof patients with chronic obstructive pulmonary disease. The limonoidsmay also be used as specific chemopreventive agents. In animal studies,results suggest that the chemotherapeutic activity of limonoids can beattributed to induction of both Phase I and Phase II detoxificationenzymes in the liver (P. Nair et al, American Journal of ClinicalNutrition, 40 (4 Suppl): 927-30, October 1984).

Another class of useful compounds that can be included in thecompositions of the present invention include the phytosterols. Thesterols occur in most plant species. Although green and yellowvegetables contain significant amounts, their seeds concentrate thesterols. Most of the research on these valuable phytonutrients has beendone on the seeds of pumpkins, yams, soy, rice and herbs. Phytosterolscompete with dietary cholesterol for uptake in the intestines. They havedemonstrated the ability to block the uptake of cholesterol (to whichthey are structurally related) and facilitate its excretion from thebody. It is well known that cholesterol is a significant risk factor incardiovascular disease. However other dietary factors also play animportant role. Research has shown the importance of other dietaryfactors in modifying the risk of cholesterol levels. A comparison of thediets of 169 Seventh Day Adventists—vegans, lacto-ovo andnon-vegetarians—with general population non-vegetarians all living inLos Angeles in the mid 1980s revealed that the ratio between dietaryplant phytosterols and cholesterol was significantly lower in SDAvegetarians as compared to non-vegetarians. The importance of this studyunderlies the fact that cholesterol, per se, is not the only marker ofrisk for cardiovascular disease and that its ratio with other modifyingdietary components may be a better measure of risk (P. Nair, et al,American Journal of Clinical Nutrition, 40 (4 Suppl): 927-30, October1984). Other investigations have revealed that phytosterols block thedevelopment of tumors in colon, breast and prostate glands. Themechanisms by which this occurs are not well understood, but we do knowthat phytosterols appear to alter cell membrane transfer in tumor growthand reduce inflammation.

Other useful phytonutrients having antioxidant activity include thephenols which comprises a large class that has been the subject ofextensive research as a disease preventive. Phenols protect plants fromoxidative damage and perform the same function for humans. Blue,blue-red and violet colorations seen in beries, grapes and purpleeggplant are due to their phenolic content. Bilberries, for example, arehigh in phenolic anthocyanidins and are red in color. The outstandingphytonutrient feature of the phenols is their ability to block specificenzymes that cause inflammation. They also modify the prostaglandinpathways and thereby protect platelets from clumping (M. G. Hertog etal., Lancet, 342: 1007-11, Oct. 23, 1993).

The flavonoids phytonutients of the above phenol subclass enhance theeffects of ascorbate-vitamin C. Flavonoids were once lumped together asvitamin P, but there are well over 1,500 of them. Among the most usefulflavonoids which can be used in the practice of the instant inventionthere are included the flavones (containing the flavonoid apigenin foundin chamomile); Flavonols (quercetin—grapefruit; rutin—buckwheat;ginkgoflavonglycosides—ginkgo); Flavanones (hsperidin—citrtis fruits;silybin—milk thisle). The biologic activities of flavonoids includeaction against allergies, inflammation, free radicals, hepatotoxins,platelet aggregation, microbes, ulcers, viruses and tumors (J. E.Kinsella et al., Food Technology, 47: 85-90, April 1993). Flavonoidsalso inhibit specific enzymes. For example, flavonoids block theangiotensin-converting enzyme (ACE) that raises blood pressure: Byblocking the suicide enzyme cyclooxygenase that breaks downprostaglandins, they prevent platelet stickiness and hence plateletaggregation. Flavonoids also protect the vascular system and strengthenthe tiny capillaries that carry oxygen and essential nutrients to allcells (J. E. Kinsella et al., Food Technology, 47: 85-90, April 1993).Additionally, flavonoids block the enzymes that produce estrogen, thusreducing the risk of estrogen-induced cancers. One way they do this isby blocking estrogen synthase, an enzyme that binds estrogen toreceptors in several organs (C. Northrup, Women's Bodies, Women'sWisdom: 305. New York; Bantam Books, 1994). Although their way of doingso is not yet fully understood, flavonoids also appear to retarddevelopment of cataracts in individuals with inborn errors in sugarmetabolism such as diabetes (R. K. Murray et al., Harper's Biochemistry,23 ed.: 196. New York; Appleton and Lange, 1994). Cataracts can be acomplication of diabetes because diabetics, unable to metabolize sugarnormally, build up damaging levels of alcohol sugars. These in turncause clouding of the lens of the eye (cataract). It is suspectedflavonoids prevent cataracts by blocking aldose-reductase (a digestiveenzyme), which can convert the sugar galactose into the potentiallyharmful form of galacticol. This select group of flavonoids deservesspecial attention. Technically known as flavonals; they providecrosslinks or bridges; that connect and strengthen the intertwinedstrands of collagen protein. Collagen is the most abundant protein inthe body, making up soft tissues, tendons, ligaments and bone matrix.Its great tensile strength depends on preservation of its crosslinks.

Anthocyanidins, being water soluble, also scavenge free radicals theyencounter in tissue fluids. This is a powerful ability especiallybeneficial for athletes and others who exercise, because heavy exercisegenerates large amounts of free radicals.

Catechins differ slightly in chemical structure from other flavonoids,but share their chemoprotective properties. The most common catechinsare gallic esters, named epicatechin (EC), epicatechin gallate (ECG),and epigallocatechin gallate (EGCG). All are found in green tea,Camellia sinensis, and are thought to be responsible for the protectivebenefits of this beverage (C. I. Xie et al., Alcohol Clin Exp Res, 18:1443-7, December 1994).

The isoflavones type of phytonutrients also included within the phenolsubclass come from beans and other legumes and are distant cousins offlavonoids. Isoflavones function much like flavonoids in that theyeffectively block enzymes that promote tumor growth. The most usefulisoflavones that can used in formulating the compositions of the presentinvention are genistein and daidzein found in soy products and the herbPueraria lobata (Kudzu). People who consume traditional diets rich insoy foods rarely experience breast, uterine and prostate cancers.Pueraria has gained popularity as an aid for those who consume alcoholbecause it appears to alter the activity of alcohol detoxificationenzymes, namely the speed at which alcohol dehydrogenase convertsalcohol into aldehydes. The result is a lowered tolerance for alcoholand reduction of the pleasure response to drinking it. Phytonutrients ofthis sulfur-containing class are present in garlic and cruciferousvegetables (i.e., cabbage, turnips and members of the mustard family).Found in cruciferous vegetables, glucosinolates are powerful activatorsof liver detoxification enzymes. They also regulate white blood cellsand cytokines (Y. Zhang et al., Proc Natl Acad Sci USA,. 91: 3147-50,Apr. 12, 1994). White blood cells are the scavengers of the immunesystem and cytokines act as messengers; coordinating the activities ofall immune cells.

Another type of flavonoids that can be used in the practice of thepresent invention is known as oligomeric proanthocyanidins (OPC's). Theyoccur in most plants, are highly bioavailable and are active in the bodyas tremendous antioxidants and free radical scavengers. OPC's arechemically classified as flavanols and are present in red wines,flowers, leaves, fruits, berries, nuts, sorghum, beans and hops withhigh concentrations in skins, barks and seeds. The most feasiblecommercial source is the seeds of grapes and the bark of the FrenchMaritime pine. Technically speaking, OPC's are really not bioflavonoids.OPC's are “flavanols” while bioflavonoids are “flavonoids.” While thechemical structure of their core molecules is the same, flavonols andflavonoids actually have more differences than similarities. OPC's arecolorless, bioflavonoids are yellow. OPC's are water soluble,bioflavonoids are relatively insoluble. Flavanols, including OPC's, arealways made up of polymers of one compound called “flavan-3-ol”, whilebioflavonoids are made up of many different compounds with a “flavane”nucleus. U.S. Pat. No. 4,698,360 whose entire contents are incorporatedby reference herein discloses the use of OPC's as antioxidants toprotect the body against the harmful biological effects of freeradicals. According to the '360 patent, OPC'S are known to cross theblood brain barrier and improve oxygenation of brain tissue, which maybe helpful in treating attention deficit syndrome. OPC's are one of themost prevalent compounds that occur in natural whole foods. Up to 25% ofOPC extract consists of monomeric precursors, the building blocks ofOPC. In grape seed extract, these precursors are known as catechins andepicatechins. Pine bark extract contains catechins and taxifolin. Everyplant species has its own unique precursor mix. Blending both grape seedand pine bark extracts combines all their monomeric precursors. The twotogether provide a broad spectrum, creating a more complete storehouseof OPC building blocks. On their own, these precursors are inactive andshort lived. However, catalyzed by the presence of OPC, they becomeactive and unfurl their own unique biological benefits rendering theblend richer in total activity.

OPC'S are probably the most versatile nutrients ever discovered. Theyare the most active free radical scavenging antioxidants known. Theyhelp protect against all degenerative conditions, such as cancer,cardiovascular diseases, stroke, allergies and aging. OPC's, orproanthocyanidins are the most powerful antioxidant free radicalscavengers known to man. They are as much as 50 times more potent thanvitamin E and 20 times more than vitamin C. OPC,s are major protectorsof collagen, the basic foundation our blood vessels and all connectivetissues such as skin, ligaments around joints and tendons.

Additional compounds that can be used also include thebio-transformation products of glucosinolates which includeisothiocyanates, dithiolthiones and sulforaphane. Each of these isprotective of specific tissues. Their actions involve blocking enzymesthat promote tumor growth, particularly in the breast, liver, colon,lung, stomach and esophagus (P. P. Tadi, Diss Abstr Int (B), 52: 4144,1992). Garlic and onions are the most potent members of this thiolsubclass, which also includes leeks, shallots and chives. The allylicsulfides in these plants are released when the plants are cut orsmashed. Once oxygen reaches the plants cells, variousbio-transformation products are formed. Each of these appears to havetissue specificity. As a group, allylic sulfides appear to possessantimutagenic and anticarcinogenic properties as well as immune andcardiovascular protection. They also appear to offer anti-growthactivity for tumors, fungi, parasites, cholesterol andplatelet/leukocyte adhesion factors. Garlic and onions, like theircruciferous relatives, can also activate liver detoxification enzymesystems. Certain allylic sulfides are known to block the activity oftoxins produced by bacteria and In viruses (P. P. Tadi, Diss Abstr Int(B), 52: 4144, 1992).

The indole subclass of phytonutrients can also be added to thecompositions of the present invention because of their interaction withvitamin C, which is not surprising since the vegetables that containindoles also contain significant amounts of vitamin C. Indole complexesbind chemical carcinogens and activate detoxification enzymes, mostly inthe gastrointestinal tract. The bio-transformation products of indolesare formed when they are acted on by stomach acid. The most activeproduct is ascorbigen, considered to be an active vitamin C metabolite(V. M. Bukhman et al, Pharmacol Ther, 60: 301-313, 1992).

An additional family of compounds which are also useful for addition toapplicants compositions include the Isoprenoids which are known toneutralize free radicals in a unique way. They have a long carbon sidechain which they use to anchor themselves into fatty membranes. Any freeradicals attempting to attach lipid (fat) membranes are quickly grabbedand passed off to other antioxidants.

A particularly preferred family of antioxidants to be combined with theother components of the present invention are the tocotrienols and thetocopherols. The tocotrienols naturally occur in grains and palm oilalong with their cousins, the tocopherols. Toco-trienols appear toinhibit breast cancer cell growth, whereas tocopherols do not exhibitthis effect. Researchers have observed that the biologic functions oftocopherols and tocotrienols appear unrelated (K. C. Hayes et al., ExpBiol Med, 202: 353-359, March 1993). Tocotrienols have been moststudied, however, for their cholesterol lowering effects.

The tocopherols are also particularly important. Vitamin E comprises agroup of natural substances known as tocopherols. These are fat soluble,closely related chemical compounds found in vegetable oils such as wheatgerm oil, soybean oil and the like. α-tocopherol has the greatestbiological activity while its homolog have vitamin E activity to alesser extent. The vitamin E can be incorporated either as the freetocopherol or as the esters i.e., d-α-tocopheryl acid succinate. Ofcourse, the racemic and nonracemic mixtures may also be used.

Lipoic acid and ubiquinone are further additives which can beincorporated in the inventive compositions. Lipoic acid and ubiquinone(coenzyme Q) are important antioxidants that work to extend the effectsof other antioxidants. Lipoic acid is known to be an efficient hydroxylradical quencher, its sulfur bond being the reactive part of themolecule. It is active on both lipids and tissue fluids. In addition tohydroxyl radicals, it scavenges peroxyl, ascorbyl and chromanoxylradicals. Because it functions in both lipid and water phases, it isprotective of both vitamin E and vitamin C. Lipoic acid also protectsSOD, catalase and glutathione, which are all important in liverdetoxification activities (R. Sumathi et al., Pharmacol Res, 27:309-318, May-June 1993). The roles of both lipoic acid and ubiquinone asantioxidants have been discovered relatively recently. Both haveimportant roles in energy production.

Another preferred antioxidant of particular interest to be used in thecompositions of the present invention is vitamin C. The vitamin C can bein the form of the acid or as sodium ascorbate or in other biologicallyacceptable mineral form such as calcium ascorbate, magnesium ascorbate,zinc ascorbate, iron ascorbate and others. Of course the ascorbic acidcan also be introduced in the form of a pro-drug e.g, as an ester whichwill hydrolyze into an inert acid and ascorbic acid i.e., ascorbylpalmitate and other fatty acid esters.

The amount of the nutriceuticals to be used in dosage forms can be anyamount that is considered safe for human consumption and approved by theacceptable guidelines promulgated by the Food and Drug Administration.For humans, typically effective amounts of histidine for use in the unitdose compositions of the present invention range from 50 mg to 32g per24 hours (0.7 to 450 mg/kg/day ); however, greater amounts may beemployed if desired. This range is based on administration to a 70 Kghuman. A preferred amount is 100 to 1500 mg per 24 hour period. Ofcourse, the amounts of each compound selected will depend on the weightof the mammal and the levels of cytoprotection, free radical scavengingeffect and antioxidant activity that are needed. One skilled in the artcan adjust the dosage forms to achieve the desired therapeutic levels.Because of histidine's favorable safety profile, large doses can also beadministered in the nutriceutical of the present invention. In humans,acute oral doses of up to 32 g/day (450 mg/kg/day ) of histidine havenot been associated with any adverse effects, and only mild, reversibleeffects were observed with acute doses up to 64 g/day.

The amount of vitamin C and other antioxidants which can be combinedwith histidine is typically in the range of 0.0001 to 10 times theamount of histidine. Of course in some cases such as with the fatsoluble antioxidants i.e., vitamin A, vitamin E and beta-carotene, theiramounts are chosen according to the National Research Council'sRecommended Dietary Allowances. Typically, for vitamin A, the adultrequirement is based on body weight and is 20 International units (IU)of retinol/Kg (one international unit equals 0.30 mcg of trans-retinolor 0.344 mcg of trans-retinol acetate. Provitamin A is required in theamount of 40 IU/kilogram (one IU of provitamin A equals 0.6 mcg ofβ-carotene). For vitamin E, the amounts will typically range from 5 to100 mg of β-tocopherol although higher amounts are possible as long asthat higher amount is not a toxic dosage. For purposes of the presentinvention 1IU=lmg of d,l α-tocopheryl acetate=0.91 mg d,lα-tocopherol=0.735 mg d α-tocopheryl acetate=0.671 mg d α-tocopherol.

The nutriceuticals of the present invention can also be combined withall of the traditional vitamin supplements. For example, histidine andall the antioxidants listed above can be formulated with the followingadditional ingredients: Vitamin A, vitamin A acetate, vitamin B₁,vitamin B₂, vitamin B₆, vitamin B₁₂, vitamin D, calcium panthotenate,niacinamide, copper, iodine, iron, magnesium, manganese, zinc, folicacid, iron, thiamine mononitrate, riboflavin, pyridoxine hydrochlorideand selenium. The above metals are typically incorporated either as thesulfates or as the carbonates.

The preferred compositions of the present invention can be prepared andadministered in a wide variety of oral, topical and parenteral dosageforms. It will be obvious to those skilled in the art that the followingdosage forms may comprise as the component, either histidine alone or incombination with other phytonutrients having antioxidant activities.

Preferably the compounds of the present invention are administeredorally, cutaneously, intramuscularly, subcutaneously, or intravenously.Oral rehydration solutions are also contemplated in the practice of thepresent invention.

For preparing nutriceutical compositions from the compounds of thepresent invention, nutriceutical acceptable carriers can be either solidor liquid. Solid form preparations include powders, tablets, pills,capsules, cachets, suppositories, and dispersible granules. A solidcarrier can be one or more substances which may also act as diluents,flavoring agents, solubilizers, lubricants suspending agents, binders,preservatives, tablet disintegrating agents, or an encapsulatingmaterial.

In powders, the carrier is a finely divided solid which is in a mixturewith the finely divided nutriceutically active component.

In tablets, the nutriceutically active component is mixed with thecarrier having the necessary binding properties in suitable proportionsand compacted in the shape and size desired.

The powders, capsules and tablets preferably contain from five or ten toabout seventy percent of the nutriceutical composition. Suitablecarriers are magnesium carbonate, magnesium stearate, talc, sugar,lactose, pectin, dextrin, starch gelatin, tragacanth, methylcellulose,sodium carboxymethylcellulose, a low melting wax, cocoa butter, and thelike. The term “preparation” is intended to include the formulation ofthe nutriceutical composition with encapsulating material as a carrierproviding a capsule in which the active component, with or without othercarriers, is surrounded by a carrier, which is thus in association withit. Similarly, cachets and lozenges are included. Tablets, powders,capsules, soft gelatin shells, pills cachets, and lozenges can be usedas solid dosage forms suitable for oral administration. Lotions,ointments, or suspensions can be used as dosage forms for topicalapplication. Anti-aging lotions, creams and onitments which also includealpha and beta hyrdoxy acids are also contemplated within the scope ofthe present invention.

For preparing suppositories, a low melting wax, such as a mixture offatty acid glycerides or cocoa butter, is first melted and the activecomponent is dispersed homogeneously therein, as by stirring. The moltenhomogeneous mixture is then poured into convenient sized molds, allowedto cool, and thereby to solidify.

Liquid for preparations include solutions, suspension, emulsions, forexample, water or water propylene glycol solutions or DMSO solutions.For parenteral injection, liquid preparations can be formulated insolution in aqueous polyethylene glycol solution or DMSO-watersolutions.

Aqueous suspensions suitable for oral use can be made by dispersing thefinely divided active component in water with viscous material, such asnatural or synthetic gums, resins, methylcellulose, sodiumcarboxymethylcellulose, and other well-known suspending agents.

Also included are solid form preparations which are intended to beconverted, shortly before use, to liquid for preparation for oraladministration. Such liquid forms include solutions, suspensions, andemulsions. These preparations may contain, in addition to the activecomponent, colorants, flavors, stabilizers, buffers, artificial andnatural sweeteners, dispersants, thickeners, solubilizing agents, andthe like. Oral rehydration solutions containing histidine are alsocontemplated by the present invention.

The nutriceutical preparation is preferably in unit dosage form. In suchform, the preparation is subdivided into unit doses containingappropriate quantities of the nutriceutically active component. The unitdosage form can be a packaged preparation, the package containingdiscrete quantities of preparation, such as packeted tablets, capsules,lotions, ointments and powders in vials or ampoules. Also, the unitdosage form can be a drink, capsule, tablet, cachet, lotion, ointment,or lozenge itself, or it can be the appropriate number of any of thesein packaged form.

When making soft gelatin shells, the shell typically consists of 20 to70 percent gelatin, generally a plasticizer and 5 to 60% by weightsorbitol. The filling of the soft gelatin capsule is liquid andincludes, apart form the antioxidant actives, a hydrophilic matrix.Typically such hydrophilic matrix comprises polyethylene glycol havingan average molecular weight of from about 200 to 1000. Furtheringredients are water and optionally thickening agents. It is preferredthat the hydrophilic matrix includes polyethylene glycol having anaverage molecular weight of from about 200 to 1000, 5 to 15% glycerol,and 5 to 15% by weight of water. The polyethylene glycol can also bemixed with propylene glycol andlor propylene carbonate.

In a preferred embodiment, there is provided a dosage unit form of thesoft gelatin shell with a filling of histidine and the otherantioxidants contemplated by the invention in combination with ahydrophilic matrix comprising polyethylene glycol having an averagemolecular weight of from about 200 to 1000, or a mixture of thepolyethylene glycol with propylene glycol, glycerol in 5 to 15% byweight, and water in 5 to 15% by weight.

As for the manufacturing, it is contemplated that standard soft shellgelatin capsule manufacturing techniques will be used to prepare thesoft-shell product. Examples of useful manufacturing techniques are theplate process, the rotary die process pioneered by R. P. Scherer, theprocess using the Norton capsule machine, and the Accogel machine andprocess developed by Lederle. Each of these processes are maturetechnologies and are all widely available to any one wishing to preparesoft gelatin capsules.

The present invention also provides a new controlled release oral tabletfor once a day administration of 500-1500 mg of D-histidine,L-histidine, a racemic mixture thereof, a non-racemic mixture thereof,and nutriceutically acceptable salts thereof, which tablet comprises ahomogeneous matrix comprising: about 3-12 weight percent ofhydroxypropylmethylcellulose having a number average molecular weight inthe range of from about 50,000 to about 150,000; about 75-97 weightpercent of D-histidine, L-histidine, a racemic mixture thereof, anon-racemic mixture thereof, and nutriceutically acceptable saltsthereof, in an amount effective for once daily oral administration; andabout 0.05 to about 4 weight percent of a pharmaceutically acceptablelubricating agent.

The tablet matrix includes a minor amount of a pharmaceuticallyacceptable lubricating agent such as magnesium stearate to aid in thetableting process. This amount will vary between about 0.05 and 4%generally, and preferably represents about 1% of the total weight of thetablet. Suitable tablet lubricants include magnesium stearate, stearicacid, calcium stearate and the like, or mixtures thereof. Magnesiumstearate is preferred.

Optionally, the tablet matrix may include minor amounts of otherpharmaceutically acceptable excipients such as colorants and glidants.Suitable colorants include, but are not limited to, FD & C Yellow #5,FD&C Yellow #6, and FD&C Blue #2, and generally represent 1% or less ofthe tablet weight. Suitable glidants include, but are not limited to,pharmaceutical grades of talc and fused silica, and generally represent7% or less of the tablet weight.

The term matrix, as used herein, refers to a uniform mixture ofD-histidine, L-histidine, a racemic mixture thereof, a non-racemicmixture thereof, and nutriceutically acceptable salts thereof,hydroxypropyl methylcellulose, a lubricating agent, and other optionallyincluded excipients. An important aspect of the present invention is thefact that the hydroxypropyl methylcellulose is uniformly dispersedthroughout the matrix to achieve uniform drug release. The matrix may bemade by any pharmaceutically acceptable technique which achieves uniformblending, including dry blending, conventional wet granulation,compression granulation, and fluid-bed granulation. Tablets can be madefrom the resulting matrix by any known tableting technique.

In accordance with the present invention, the amount of D-histidine,L-histidine, a racemic mixture thereof, a non-racemic mixture thereof,and nutriceutically acceptable salts thereof, that is incorporated in atablet may range between about 500 and about 1500 mg. The tablet of thepresent invention provides a release period suitable for once-dailydosing, i.e. once within a 24 hour period. Generally, D-histidine,L-histidine, a racemic mixture thereof, a non-racemic mixture thereof,and nutriceutically acceptable salts thereof, are administered at levelsof 500, 750, 1000 or 1500 mg/day.

The hydroxypropylmethylcellulose utilized in the present invention is awater soluble cellulose ether, and is commercially available in variousgrades under the tradenames Methocel Premium K4M, A4M, E4M and F4M (DowChemical Co., U.S.A.) which are 4000 cps viscosity polymers havingnumber average molecular weights in the range of 85,000-95,000, andMethocel K15M, a 15,000 cps viscosity polymer having a number averagemolecular weight in the range of 120,000-130,000. Other suitablepolymers include Metolose SM, 60SH, 65SH, and 90SH, viscosity grades4000, 8000, 15,000 and 30,000 available from Shin-Etsu Ltd., Japan.Specific preferred hydroxypropyl methylcelluloses are Methocel K4MPremium and Methocel K15M Premium.

The physicochemical properties of these polymers vary over a wide range.Preferred embodiments of this invention utilize premium grade polymersof a single viscosity type having number average molecular weights inthe range of about 50,000-150,000. However, a mixture of two or moregrades or viscosity types resulting in number average molecular weightsin the range of about 50,000-150,000 can be used.

The number average molecular weight of the hydroxypropyl methylcellulosewhich is used in the tablet matrix substantially influences the releaseprofile which is obtained. The number average molecular weight (Mn) isthe sum of the individual molecular weights of a representative samplepopulation of molecules divided by the number of molecules in thatsample, and is calculated from the limiting osmotic pressure of thesolvent as the concentration of the hydroxypropyl methylcelluloseapproaches zero. The hydroxypropyl methylcellulose must have a numberaverage molecular weight in the range of from about 50,000 to about150,000, preferably from about 125,000 to about 135,000. When thepolymer has a number average molecular weight of 125,000-135,000, itconstitutes preferably about 3-12 weight percent of a 750 to 1000 mgL-Histidine controlled release tablet, or about 7 weight percent of a500 to 750 mg L-Histidine controlled release tablet. A second preferredrange of number average molecular weight of the hydroxypropylmethylcellulose is about 85,000 to about 95,000. When the polymer has anumber average molecular within this range, it constitutes preferablyabout 7-9 weight percent of the controlled release L-Histidine tablet.

Optionally, the controlled release tablets may be coated with anyconventional or non-conventional coating which is pharmaceuticallyacceptable. Generally, the coatings will be applied for such purposes asproduct identification, printability of the tablet, light protection,aesthetic appearance and patient compliance, and will not effect therelease rate profile of the tablet. Conventional coating formulationswhich may be used include aqueous and solvent-based polymeric, gum andcellulose-based films, as well as sugar coatings. Numerouspharmaceutically acceptable coating formulations have been developed bythe pharmaceutical industry and are well known to the pharmaceuticalscientist.

Exemplary of typical film coating materials are formulations of suchfilm forming agents as polyvinylpyrrolidone, acrylic polymers andcopolymers, natural gums and resins, low molecular weight hydroxypropylmethylcellulose, hydroxypropyl cellulose, ethyl cellulose andmethylcellulose. These formulations may include plasticizing agents suchas acetylated monoglycerin, castor oil, mineral oil, glycerol, diethylphthalate, triethylcitrate, triacetin, glycerin, propylene glycol,polyethylene glycol, and the like, as well as surface active agents suchas polysorbates, colorants, and opacifiers such as titanium dioxide andtalc.

A preferred coating formulation for the tablets of this inventioncomprises a mixture of low molecular weight hydroxypropylmethylcellulose and polyethylene glycol. Such coatings are readilyformulated by standard means well known to the pharmaceutical chemist,and are also commercially available. Exemplary of a commerciallyavailable coating material suitable for application to the tablets ofthis invention is OpadryRTM., a low molecular weight hydroxypropylmethylcellulose/polyethylene glycol material available from Colorcon.

The controlled release tablets of the present invention providenutriceutically effective blood levels of L-Histidine for at least 12 to24 hours, and are thus suitable for once-daily administration.Fluctuations in blood levels during multi-dose therapeutic regimens areminimized by the tablets of the present invention.

The examples described below are presented for illustration purposesonly and are not intended to limit the scope of the invention of thisapplication which is defined in the claims. The present invention isfurther explained more specifically with reference to the followingformulation examples without limit thereto. The parts and percentages inthe formulation Examples are represented by weight:

EXAMPLE I

The following example illustrates a histidine formulation which can bemade into a capsule:

COMPONENT AMOUNT Histidine 250 mg Lactose q.s to selected size

The active ingredients are triturated and q.s. with lactose to selectedcapsules size.

EXAMPLE II

COMPONENTS AMOUNT Histidine 125 mg Vitamin C 125 mg Vitamin E 75 I.U.Anhydrous glycerol 35 mg Polyethylene glycol 400 315 mg

The above components are thoroughly mixed under a nitrogen atmosphere,and are then encapsulated in conventional soft gelatin shells.

EXAMPLE III

COMPONENTS AMOUNT Histidine 350 mg Vitamin C 145 mg β-Carotene 5000 I.U.Anhydrous glycerol 35 mg Polyethylene glycol 400 315 mg

The above components are thoroughly mixed under a nitrogen atmosphere,and are then encapsulated in conventional soft gelatin shells.

EXAMPLE IV

COMPONENTS AMOUNT Histidine 400 mg Vitamin E 50 I.U. β-Carotene 4500I.U. Anhydrous glycerol 35 mg Polyethylene glycol 400 315 mg

The above components are thoroughly mixed under a nitrogen atmosphere,and are then encapsulated in conventional soft gelatin shells.

EXAMPLE V

COMPONENTS AMOUNT Histidine 285 mg Vitamin A 1700 I.U. β-Carotene 5000I.U. Anhydrous glycerol 35 mg Polyethylene glycol 400 315 mg

The above components are thoroughly mixed under a nitrogen atmosphere,and are then encapsulated in conventional soft gelatin shells.

EXAMPLE VI

COMPONENTS AMOUNT Histidine 250 mg Vitamin C 245 mg β-Carotene 2500 I.U.Anhydrous glycerol 35 mg Polyethylene glycol 400 315 mg

The above components are thoroughly mixed under a nitrogen atmosphere,and are then encapsulated in conventional soft gelatin shells.

EXAMPLE VII

COMPONENTS AMOUNT Histidine 300 mg Vitamin A 850 I.U. β-Carotene 5000I.U. Vitamin C 200 mg Vitamin E 70 I.U. Anhydrous glycerol 35 mgPolyethylene glycol 400 315 mg

The above components are thoroughly mixed under a nitrogen atmosphere,and are then encapsulated in conventional soft gelatin shells.

EXAMPLE VIII

COMPONENTS AMOUNT Histidine 200 mg Vitamin A 2000 I.U. β-Carotene 3000I.U. Magnesium sulfate 100 mg Vitamin C 175 mg Anhydrous glycerol 35 mgPolyethylene glycol 400 315 mg

The above components are thoroughly mixed under a nitrogen atmosphere,and are then encapsulated in conventional soft gelatin shells.

EXAMPLE IX

Hard gelatin capsules are prepared using the following ingredients:

COMPONENT mg/Capsule Histidine 250 Starch dried 200 Magnesium Stearate10

The above ingredients are mixed and filled into hard gelatin capsules in460 mg quantities.

EXAMPLE X

A tablet formula is prepared using the ingredients below:

COMPONENT mg/Tablet Histidine 150 Vitamin C 100 Microcrystallinecellulose 400 Fumed Silicon Dioxide 10 Stearic Acid 5

The components are blended and compressed to form tablets each weighing665 mg.

EXAMPLE XI

Tablets each containing 60 mg of active ingredients are made up asfollows:

COMPONENT mg/Tablet Histidine 20 Vitamin C 20 Vitamin E in dry carrier20 Starch 45 Microcrystalline Cellulose 35 Polyvinylpyrrolidone (as 10%4.0 solution in water) Sodium Carboxymethyl starch 4.5 Magnesiumstearate 0.5 Talc 1.0

The nutriceutically active ingredients ,starch and cellulose are passedthrough a No. 45 mesh U.S. sieve and mixed thoroughly. The solution ofpolyvinylpyrrolidone is mixed with the resultant powders which are thenpassed through a No. 14 mesh U.S. sieve. The granules so produced aredried at 50-60° C. and passed through a No. 18 mesh U.S. sieve. Thesodium carboxymethyl starch, magnesium stearate and talc, previouslypassed through a No. 60 mesh U.S. sieve, are then added to the granuleswhich, after mixing, are compressed by a tablet machine to yield tabletseach weighing 150 mg.

EXAMPLE XII

Capsules each containing 80 mg of nutriceuticals are made as follows:

COMPONENTS mg/Capsule L-Histidine 30 mg Vitamin C 35 mg Vitamin E 14 mgβ-Carotene  1 mg Starch 59 mg Microcrystalline Cellulose 59 mg MagnesiumStearate  2 mg

The nutriceutical active ingredients, cellulose, starch and magnesiumstearate are blended, passed through a No. 45 mesh U.S. sieve, andfilled into hard gelatin capsules in 200 mg quantities.

EXAMPLE XIII

Suspensions each containing 50 mg of histidine per 5 ml dose are made asfollows:

COMPONENT AMOUNT L-Histidine 50 mg Sodium Carboxymethyl cellulose 50 mgSyrup 1.25 ml Benzoic Acid solution 0.10 ml Flavor q.v. Color q.vPurified water to 5 ml

The histidine is passed through a No. 45 mesh U.S. sieve and mixed withthe sodium carboxymethylcellulose and syrup to form a smooth paste. Thebenzoic acid solution, flavor and color are diluted with some of thewater and added, with stirring. Sufficient water is then added toproduce the required volume.

EXAMPLE XIV

Suspensions each containing 25 mg of histidine and 25 mg of vitamin Cper 5 ml dose are made as follows:

COMPONENT AMOUNT L-Histidine 25 mg Vitamin C 25 mg Sodium Carboxymethylcellulose 50 mg Syrup 1.25 ml Benzoic Acid solution 0.10 ml Flavor q.v.Color q.v Purified water to 5 ml

The histidine and vitamin C are passed through a No. 45 mesh U.S. sieveand mixed with the sodium carboxymethylcellulose and syrup to form asmooth paste. The benzoic acid solution, flavor and color are dilutedwith some of the water and added, with stirring. Sufficient water isthen added to produce the required volume.

EXAMPLE XV

Capsules each containing 150 mg of nutriceuticals are made as follows:

COMPONENT mg/Capsule Histidine 50 mg Vitamin C 50 mg Vitamin E 45 mgVitamin A  4 mg β-Carotene  1 mg Starch 164 mg  MicrocrystallineCellulose 164 mg  Magnesium Stearate 22 mg

The active ingredient, cellulose, starch and magnesium stearate areblended, passed through a No. 45 mesh U.S. sieve, and filled into hardgelatin capsules in 500 mg quantities.

EXAMPLE XVI

Example XV is repeated except one microgram of selenate is added to theformulation.

EXAMPLE XVII SUSTAINED RELEASE COMPOSITION CONTAINING HISTIDINE

350 grams of histidine crystals, all having a size of between 20 and 70mesh, are placed in a six inch air suspension coating column (Wurstercolumn of manufacture by Glatt, West German) and coated with a mixtureof 184 ml polymer solution in chloroform which contained 15 gmethylcellulose [“Ethocel N-10” (Dow)] and 3.7 gm hydroxypropyl cellulose[“Klucel LF” (Hercules)] and 46 ml of methanol. The coating solution issprayed at 2.5 bar pressure with the liquid feed rate of 30 ml/minute.The inlet air temperature is about 60° C. After completion of the feedof the coating, the quickly dried polymerically coated histidinecrystals are recovered from the bottom of the air suspension coatingcolumn.

EXAMPLE XVIII SUSTAINED RELEASE COMPOSITION CONTAINING HISTIDINE ANDVITAMIN C

350 grams of histidine crystals and 350 grams of vitamin C crystals, allhaving a size of between 20 and 70 mesh, are placed in a six inch airsuspension coating column (Wurster column of manufacture by Glatt, WestGerman) and coated with a mixture of 368 ml polymer solution inchloroform which contained 30 gm ethylcellulose [“Ethocel N-10” (Dow)]and 7.4 gm hydroxypropyl cellulose [“Klucel LF” (Hercules)] and 92 ml ofmethanol. The coating solution is sprayed at 2.5 bar pressure with theliquid feed rate of 60 ml/minute. The inlet air temperature is about 60°C. After completion of the feed of the coating, the quickly driedpolymerically coated histidine and vitamin C crystals are recovered fromthe bottom of the air suspension coating column.

EXAMPLE XIX

Sustained release tablets containing L-Histidine are made as follows:3500 gms of L-histidine and 180 gms Methocel K15M were well blended, andthen granulated with about 1000 ml of purified water. The granulationwas tray dried in a 50 degree. C. oven for 16 hours, passed at slowspeed through a hammer mill fitted with an 18 gauge screen, and thenthoroughly mixed with 39 gms of magnesium stearate. The resultinghomogeneous matrix material was compressed into tablets of about 800 mgin weight and size.

The following represent additional formulation examples:

EXAMPLE XX

COMPONENTS AMOUNT Histidine 250 mg Vitamin C 245 mg lycopene 2500 I.U.Anhydrous glycerol 35 mg Polyethylene glycol 400 315 mg

The above components are thoroughly mixed under a nitrogen atmosphere,and are then encapsulated in conventional soft gelatin shells.

EXAMPLE XXI

COMPONENTS AMOUNT Histidine 250 mg lycopene 2500 I.U. Anhydrous glycerol35 mg Polyethylene glycol 400 315 mg

The above components are thoroughly mixed under a nitrogen atmosphere,and are then encapsulated in conventional soft gelatin shells.

EXAMPLE XXII

COMPONENTS AMOUNT Histidine 250 mg Lipoic acid 0.1 mg Anhydrous glycerol35 mg Polyethylene glycol 400 315 mg

The above components are thoroughly mixed under a nitrogen atmosphere,and are then encapsulated in conventional soft gelatin shells.

Various other examples and modifications or adaptations of the foregoingExamples can be devised by a person skilled in the art after reactingthe foregoing specification and the appended claims without departingfrom the spirit and scope of the invention. All such further Examples,modifications and adaptations are included within the scope of theinvention.

It will be appreciated by those versed in the art, that the presentinvention makes available novel and useful nutriceutical compositionshaving cytoprotective effects in several administration forms includingsustained release. Also, it will be understood by those with knowledgein the nutriceutical art, that many embodiments of this invention may bemade without departing from the spirit and scope of the invention, andthe invention is not to be construed as limited, as it embraces allequivalents therein.

What is claimed is:
 1. A dietary supplement comprising: (a) an effectiveamount of a synthetic or naturally occurring phytonutrient selected fromthe group consisting of canthanaxin, limonene and vitamin A; (b) acytoprotective effective amount of histidine wherein said histidine isselected from the group consisting of at least one of D-histidine,L-histidine, a racemic mixture thereof, a non-racemic mixture thereof,and nutriceutically acceptable salts thereof, and a nutriceuticallyinert carrier selected from the group consisting of microcrystallinecellulose, polyethylene glycol, sodium carboxymethyl cellulose andhydroxypropyl methylcellulose.
 2. A nutricoutical histidine compositioncomprising: (a) an effective cyto-protective amount of histidine whereinsaid histidine is selected from the group consisting of at least one ofD-histidine, L-histidine, a racemic mixture thereof, a non-racemicmixture thereof, and nutriceutically acceptable salts thereof; (b) aneffective amount of one or more of a phytonutrient having antioxidantactivity selected from the group consisting of canthaxantin, limonene,vitamin A and mixtures thereof; and (c) a nutrceutically inert carrierselected from the group consisting of microcrystalline cellulose,polyethylene glycol, sodium carboxymethyl cellulose and hydroxypropylmethylcellulose.
 3. A method of providing nutrition to a mammal in needthereof, which method comprises administering to said mammal acomposition comprising: (a) an effective amount of a cyto-protectiveagent selected from the group consisting of at least one of D-histidine,L-histidine, a racemic mixture thereof, a non-racemic mixture thereof,and nutriceulically acceptable salts thereof; and (b) an effectiveamount of at least one of a phytonutrient having antioxidant activityselected from the group consisting of canthaxantin, limonene, vitamin Aand mixtures thereof.